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Distribution and serotonin-induced activation of vacuolar-type H+-ATPase in the salivary glands of the blowfly Calliphora vicina
1 Institut für Biochemie und Biologie, Zoophysiologie, Universität
Potsdam, Lennéstr. 7a, D-14471 Potsdam, Germany
2 Carl Zeiss Jena GmbH, Advanced Imaging Microscopy, Carl-Zeiss-Promenade
10, D-07745 Jena, Germany
* Author for correspondence (e-mail: obaumann{at}rz.uni-potsdam.de)
Accepted 14 March 2003
Secretory activity in blowfly salivary glands is activated by the hormone serotonin. We have investigated the distribution and activity of two cation pumps that are possibly involved with transepithelial ion transport, i.e. Na+/K+-ATPase and vacuolar-type H+-ATPase (V-ATPase). By immunofluorescence labelling of secretory cells, Na+/K+-ATPase was localized on the basolateral plasma membrane and V-ATPase on the highly folded apical membrane. Activities of both ATPases were probed in salivary gland homogenates by applying specific inhibitors for these ion pumps, namely ouabain and bafilomycin A1. In control glands, bafilomycin-A1-sensitive V-ATPase activity and ouabain-sensitive Na+/K+-ATPase activity accounted for 36% and 19%, respectively, of the total ATPase activity. V-ATPase activity increased approximately twofold after stimulation with serotonin, whereas Na+/K+-ATPase activity was not significantly affected. Biochemical assays provided evidence that the serotonin-induced activation of V-ATPase activity was accompanied by a recruitment of peripheral V1 subunits from the cytosol to the plasma membrane, indicative of the assembly of V0V1 holoenzymes.
These data show that a V-ATPase located in the apical plasma membranes of the secretory cells is a component of the apical `potassium pump' that has been identified previously by physiological approaches. The V-ATPase energizes the apical membrane and provides the primary driving force for fuelling a putative K+/nH+ antiporter and, thus, for fluid secretion. Serotonin-induced assembly of V0V1 holoenzymes might constitute a regulatory mechanism for the control of pump activity.
Key words: vacuolar ATPase, V-ATPase, portasome, bafilomycin, Na+/K+-ATPase, serotonin, 5-hydroxytryptamine, regulation, assembly, immunocytochemistry, insect, blowfly, Calliphora vicina.
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